Les auteurs remercient le technicien dentaire Nicola Lanfranconi pour la production du modèle de référence maître et la Société Alicona pour leur soutien continu à l&#39;amélioration du logiciel de numérisation.

1. Préparation des échantillons / base <ol><li> Appliquer un fond plat autour de l&#39;échantillon. Placer l&#39;échantillon sur la table de balayage. Orienter la surface de mastication par rapport au plan horizontal (figure 1). </li></ol> 2. Analyse de logiciel <ol><li> Démarrez le programme puis le module de mesure de laboratoire (Figure 2) </li><li> Positionner l&#39;échantillon au centre de la table de numérisation. </li><li> Choisissez la lentille de grossissement correct. Pour les grands objets, comme des analyses de arcade complète, utilisez l&#39;objectif 5X. </li><li> Déplacez l&#39;optique de balayage à l&#39;aide de la souris 3D jusqu&#39;à ce que la surface de l&#39;échantillon est affiché sur la fenêtre d&#39;affichage en direct (figure 3). </li><li> Utilisez la commande de sonde pour régler l&#39;exposition et le contraste pour obtenir les paramètres de numérisation optimales. Pour les surfaces en métal, utiliser une exposition entre 400 et 800 ps et un contraste entre 0,3 et 0,8 (figure 4). </li><li> Vérifiez la qualité de l&#39;image en cochant le bouton&quot;Afficher la qualité d&#39;image dans l&#39;aperçu en direct&quot;. </li><li> Définissez les paramètres corrects dans la section &quot;Mesure de la commande&quot;. Le type de mesure est &quot;3D Dataset&quot;; ImageField Type est &quot;général ImageField&quot; (Figure 5). </li><li> Cliquez sur &quot;Nouveau ImageField&quot;. </li><li> Définir le volume de numérisation. Le logiciel a besoin des coordonnées de délimitation du volume de mesure. <ol><li> Déplacez le spécimen au plus haut niveau et le plus bas de numérisation et cliquez sur &quot;Ajouter Position&quot; à deux points. La valeur &quot;Range Z&quot; représente la hauteur réelle du volume de balayage (Figure 6). </li><li> Déplacer l&#39;échantillon aux boundings XY du volume de scan. Cliquez sur &quot;Ajouter Position&quot; pour définir l&#39;axe X et l&#39;axe Y longueurs. La dimension du volume de balayage est affiché dans l&#39;onglet &quot;Information&quot; et devrait dépasser les dimensions de l&#39;échantillon de 1 cm dans l&#39;axe X et l&#39;axe Y. </li></ol></li><li> Vérifiez le nombre de «points». Le logiciel est capable de scanner 100 millions de points de surface dans un &quot;ImageField&quot; balayage. Le nombre réelde points dépasse cette limite. &quot;Décimer&quot; le numéro du point de mesure par un &quot;sous-échantillonnage latéral&quot; de la taille de point. <ol><li> Cliquez sur &quot;Paramètres avancés&quot; (figure 7). </li><li> Déplacez le curseur &quot;latéral sous-échantillonnage&quot; à la droite jusqu&#39;à ce que le nombre de «points» est réduite en dessous de 100 M (figure 7). Le sous-échantillonnage réduit la résolution latérale des points de la surface, ce qui entraîne une plus grande taille de pixel de l&#39;objet balayé. Pour assurer des résultats optimaux de numérisation, la résolution verticale est augmentée. </li></ol></li><li> Cliquez sur &quot;Démarrer la mesure&quot;. Cela va démarrer le &quot;Mode de Prévisualisation&quot;. Le logiciel effectue un pré-balayage avec les dimensions X et Y, l&#39;axe sélectionné. </li><li> Après le pré-scan est terminé, sélectionnez la région d&#39;intérêt. Cela contribue à réduire la taille du fichier et le temps de balayage (Figure 8). </li><li> Sélectionner toutes les régions du prescan sauf l&#39;échantillon et deux champs de mesure autour de l&#39;échantillon, contenant la base plat (figure 9). </li><li> Cliquez sur &quot;Démarrer&quot; pour lancer la numérisation. </li><li> Contrôler le balayage dans la &quot;Visionneuse&quot; avec la souris et appuyez sur le bouton gauche de la souris (figure 10). </li><li> Fermez la fenêtre et cliquez sur &quot;Afficher Pseudo Color Seulement&quot;. </li><li> Cliquez sur &quot;Paramètres&quot; et &quot;pseudo Coloriage&quot;, et choisissez &quot;répétabilité&quot;. </li><li> Réglez le &quot;Max.&quot; la valeur de 0,2, puis cliquez sur &quot;Appliquer la plage&quot;. </li><li> Contrôler la répétabilité, il doit être égal pour les zones avec la même inclinaison et matériel. En particulier, la base autour de l&#39;échantillon doit afficher une répétabilité homogène (Figure 11). </li><li> Cliquez sur &quot;base de données&quot; et enregistrer la numérisation vers le dossier approprié. </li><li> Exporter l&#39;analyse de différents formats de fichiers, si nécessaire. Cliquez sur &quot;File/Export/3D données Comme / ....&quot; Le protocole peut être mis en pause à ce moment-là et a continué plus tard. </li></ol> 3. Analyse de différence <ol><li> Utilisez la &quot;3D-Editor&quot; pour couper la base. Cette zone ne sera pas utilisée pour la différenceanalyse. </li><li> Pour comparer et d&#39;analyser deux balayages, démarrez le logiciel &quot;Différence de mesure» (figure 12). </li><li> Choisissez le second modèle à comparer. </li><li> Cliquez sur &quot;Alignement automatique rugueux&quot; pour effectuer un premier match des modèles. </li><li> Cliquez sur &quot;Réglage manuel&quot; et aligner les modèles de tourner et déplacer jusqu&#39;à ce qu&#39;ils soient dans le même sens (figure 13). </li><li> Cliquez sur &quot;Alignement automatique&quot; et &quot;Appliquer&quot; pour commencer le meilleur algorithme d&#39;ajustement pour le modèle optimal correspondant. </li><li> Cliquez sur &quot;différences&quot; pour voir la carte de différence entre les deux modèles assortis. </li><li> Sélectionnez une plage de couleur appropriée pour afficher les écarts (figure 14). Enregistrez le dessin d&#39;écart visuel comme une capture d&#39;écran pour l&#39;analyse. </li></ol> Cliquez sur &quot;Statistiques&quot; pour afficher les valeurs statistiques des différences. Choisissez une taille de classe um et enregistrer les données de l&#39;histogramme dans un fichier texte pour la comparaison statistique (Figure 15).

<ol>
	<li>Wettstein, F., Sailer, I., Roos, M., H&#228;mmerle, C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clinical+study+of+the+internal+gaps+of+zirconia+and+metal+frameworks+for+fixed+partial+dentures.">Clinical study of the internal gaps of zirconia and metal frameworks for fixed partial dentures.</a> Eur. J. Oral Sci. 116 (3), 272-279 (2008).</li><li>Persson, A. S., Oden, A., Andersson, M., Sandborgh-Englund, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Digitization+of+simulated+clinical+dental+impressions:+virtual+three-dimensional+analysis+of+exactness.">Digitization of simulated clinical dental impressions: virtual three-dimensional analysis of exactness.</a> Dent. Mater. 25 (7), 929-936 (2009).</li><li>Del'Acqua, M. A., Arioli-Filho, J. N., Compagnoni, M. A., Mollo, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=de+A+Accuracy+of+impression+and+pouring+techniques+for+an+implant-supported+prosthesis.">de A Accuracy of impression and pouring techniques for an implant-supported prosthesis.</a> Int. J. Oral Maxillofac. Implants. 23 (2), 226-236 (2008).</li><li>Schneider, D., Grunder, U., Ender, A., H&#228;mmerle, C. H. F., Jung, R. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Volume+gain+and+stability+of+peri-implant+tissue+following+bone+and+soft+tissue+augmentation:+1-year+results+from+a+prospective+cohort+study.">Volume gain and stability of peri-implant tissue following bone and soft tissue augmentation: 1-year results from a prospective cohort study.</a> Clinical Oral Implants Research. 22 (1), 28-37 (2011).</li><li>Windisch, S. I., Jung, R. E., Sailer, I., Studer, S. P., Ender, A., H&#228;mmerle, C. H. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+optical+method+to+evaluate+three-dimensional+volume+changes+of+alveolar+contours:+a+methodological+in+vitro+study.">A new optical method to evaluate three-dimensional volume changes of alveolar contours: a methodological in vitro study.</a> Clinical Oral Implants Research. 18 (5), 545-551 (2007).</li><li>Caputi, S., Varvara, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dimensional+accuracy+of+resultant+casts+made+by+a+monophase,+one-step+and+two-step,+and+a+novel+two-step+putty/light-body+impression+technique:+an+in+vitro.">Dimensional accuracy of resultant casts made by a monophase, one-step and two-step, and a novel two-step putty/light-body impression technique: an in vitro.</a> 99 (4), 274-281 (2008).</li><li>Hoyos, A., Soderholm, K. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Influence+of+tray+rigidity+and+impression+technique+on+accuracy+of+polyvinyl+siloxane+impressions.">Influence of tray rigidity and impression technique on accuracy of polyvinyl siloxane impressions.</a> Int. J. Prosthodont. 24 (1), 49-54 (2011).</li><li>Luthardt, R. G., Kuhmstedt, P., Walter, M. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+method+for+the+computer-aided+evaluation+of+three-dimensional+changes+in+gypsum+materials.">A new method for the computer-aided evaluation of three-dimensional changes in gypsum materials.</a> Dent. Mater. 19 (1), 19-24 (2003).</li><li>Mehl, A., Ender, A., M&#246;rmann, W., Attin, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Accuracy+testing+of+a+new+intraoral+3D+camera.">Accuracy testing of a new intraoral 3D camera.</a> International Journal of Computerized Dentistry. 12 (1), 11-28 (2009).</li><li>Fickl, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dimensional+changes+of+the+ridge+contour+after+socket+preservation+and+buccal+overbuilding:+an+animal+study.">Dimensional changes of the ridge contour after socket preservation and buccal overbuilding: an animal study.</a> J. Clin. Periodontol. 36 (5), 442-448 (2009).</li><li>Vlaar, S. T., vander Zel, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Accuracy+of+dental+digitizers.">Accuracy of dental digitizers.</a> Int. Dent. J. 56 (5), 301-309 (2006).</li><li>Ender, A., Mehl, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Accuracy+of+complete-arch+dental+impressions:+a+new+method+of+measuring+trueness+and+precision.">Accuracy of complete-arch dental impressions: a new method of measuring trueness and precision.</a> 109 (2), 121-128 (2013).</li><li>Ender, A., Mehl, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Influence+of+scanning+strategies+on+the+accuracy+of+digital+intraoral+scanning+systems.">Influence of scanning strategies on the accuracy of digital intraoral scanning systems.</a> Int. J. Comput. Dent. 16 (1), 11-21 (2013).</li><li>Ender, A., Mehl, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Full+arch+scans:+conventional+versus+digital+impressions--an+in-vitro+study.">Full arch scans: conventional versus digital impressions--an in-vitro study.</a> Int. J. Comput. Dent. 14 (1), 11-21 (2011).</li><li>Meer, W. J., Andriessen, F. S., Wismeijer, D., Ren, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Application+of+intra-oral+dental+scanners+in+the+digital+workflow+of+implantology.">Application of intra-oral dental scanners in the digital workflow of implantology.</a> PLoS One. 7 (8), (2012).</li></ol>

Les auteurs déclarent qu&#39;ils n&#39;ont aucun intérêt financier concurrents ou d&#39;autres conflits d&#39;intérêts.

La précision est une exigence fondamentale en médecine dentaire. Le scanner de référence est capable de numériser objets petits et grands avec une grande justesse et précision. Avec la méthode de numérisation optimale, même sur des surfaces dentaires détaillées morphologiques individuels peuvent être analysés avec une haute résolution et la répétabilité. Avec les différents niveaux du scanner de grossissement, les structures morphologiques macro et micro peuvent être acquises. Il est possible de balay...

La précision est d&#39;un intérêt majeur dans de nombreux domaines de la médecine dentaire. Remplacement des tissus durs dentaires a besoin d&#39;une prothèse de montage exacte pour assurer le bon fonctionnement et éviter de détruire davantage la structure restante de la dent 1,2. Prothèses partielles fixes et prothèse totale sont particulièrement critiques pour le montage exact des structures de soutien comme des dents ou des implants 3 préparés. C&#39;est la raison pour laquelle une reproduction très précise est nécessaire, en particulier dans le domaine des empreintes dentaires et les flux de travail de laboratoire dentaire. Toutefois, d&#39;autres domaines de traitement dentaire bénéficient également d&#39;un résultat vrai et précis métrique, pour vérifier le succès du traitement et évaluer de nouvelles stratégies de traitement, par exemple l&#39;augmentation des tissus mous et durs, l&#39;érosion et à l&#39;abrasion, la surveillance des traitements parodontaux, et les traitements orthodontiques 4,5. Dans beaucoup de ces domaines, des procédures de validation actuelles sont des mesures de distance linéaire avec étriers ou des microscopes 6,7. Ces méthods sont limités à seulement quelques points de mesure et des informations limitées de trois dimensions (3D) des modifications de la zone de test. Méthodes de mesure les plus récents incluent la capture optique ou radiographique de toute la surface du test objet 8,9. Ici, la surface ou le volume entier est mesuré et affiché comme un objet 3D sur l&#39;écran d&#39;ordinateur. Mesures linéaires sont possibles, ainsi que des superpositions de modèles de différentes époques de balayage. Avec cette superposition, une évaluation des modifications de surface à chaque point de balayage est possible. Ceci permet la possibilité de surveiller une zone spécifique ou de l&#39;affichage des déformations dans les trois axes de coordonnées. En outre, les changements volumétriques peuvent être mesurées 10. Le point limite à ces nouvelles méthodes est la précision du dispositif de balayage, utilisé pour capturer l&#39;objet de test. Aucune des modifications au sein de l&#39;exactitude du dispositif de balayage de référence peut être divisée en des changements de l&#39;objet d&#39;essai ou des erreurs d&#39;analyse. Précision de balayage est souvent une valeur donnée par le fabricantcant dérivé de la numérisation de petits objets calibrés 11. Cette erreur d&#39;analyse minimale est différente lors de la numérisation de gros objets comme une arcade dentaire. Précision compose de justesse et de précision. L&#39;exactitude est la déviation de l&#39;objet balayé de sa géométrie réelle. La précision est l&#39;écart entre les balayages répétés (ISO 5725-1). Dans cette étude, un nouveau scanner optique de référence, sur la base de la variation technique de balayage de mise au point, a été introduit pour analyser les échantillons de dent à des modèles complets de voûte avec la plus grande exactitude. Ce scanner de référence a été utilisé comme une base pour plusieurs études, comparant la précision d&#39;empreinte dentaire de techniques classiques et numériques 12-14 et pour des projets concrets concernant l&#39;occlusion dentaire et à l&#39;abrasion des matériaux dentaires. L&#39;objectif de cette étude était de fournir des informations de base de la précision du scanner de référence et des possibilités d&#39;utiliser ce dispositif dans le domaine de la recherche dentaire.

<table border="0" cellpadding="0" cellspacing="0" width="669">
	<tbody>
		<tr>
			<td>Reference model</td>
			<td></td>
			<td></td>
			<td>individual non-precious metal model, derived from a patient impression</td>
		</tr>
		<tr>
			<td>Araldit repair</td>
			<td>Huntsmen Advanced Material, Basel, Switzerland</td>
			<td></td>
			<td>used for making the base of the reference model</td>
		</tr>
		<tr>
			<td>CamBase</td>
			<td>Dentona, Dortmund, Germany</td>
			<td></td>
			<td>Type IV dental ston for pouring conventional impressions</td>
		</tr>
		<tr>
			<td>Identium</td>
			<td>Kettenbach, Eschenburg, Germany</td>
			<td></td>
			<td>Vinylsiloxanether impression material for conventional impression</td>
		</tr>
		<tr>
			<td>inEOS model holder</td>
			<td>Sirona Dental Systems, Bensheim, Germany</td>
			<td></td>
			<td>used for fixing stone models at the reference scanner</td>
		</tr>
		<tr>
			<td>Accutrans</td>
			<td>Coltene Whaledent, Altst&#228;tten, Switzerland</td>
			<td></td>
			<td>used for making the base of thestone models</td>
		</tr>
		<tr>
			<td>President putty</td>
			<td>Coltene Whaledent, Altst&#228;tten, Switzerland</td>
			<td></td>
			<td>mix with accutrans for betterstability of the base</td>
		</tr>
		<tr>
			<td>Alicona Infinite Focus</td>
			<td>Alicona Imaging, Graz, Austria</td>
			<td></td>
			<td>Reference scanner&#160;</td>
		</tr>
	</tbody>
</table>

accuracy in dental medicine, a new way to measure trueness and precision

scanners de référence sont utilisées en médecine dentaire pour vérifier un grand nombre de procédures. L&#39;intérêt principal est de vérifier les méthodes d&#39;impression car ils servent de base pour les restaurations dentaires. La limitation de courant de nombreux scanners de référence est l&#39;absence de balayage de précision de gros objets comme des arcades dentaires complets, ou la possibilité limitée pour évaluer la surface des dents détaillées. Un nouveau scanner de référence, basé sur la variation technique de balayage de mise au point, a été évaluée en ce qui concerne la plus grande précision locale et générale. Un protocole d&#39;analyse spécifique a été testée pour balayer la surface de la dent d&#39;origine à partir des empreintes dentaires. En outre, des matériaux modèles différents ont été vérifiés. Les résultats ont montré une précision de numérisation haute du scanner de référence avec un écart moyen de 5,3 ± 1,1 um pour la justesse et de 1,6 ± 0,6 um pour la précision en cas de scans arcade complète. Méthodes d&#39;empreintes dentaires actuelles ont montré beaucoup plus élevé écarts (justesse: 20,4 ± 2,2 um, précision: 12,5 ± 2,5 um) than la Précision d&#39;analyse interne du scanner de référence. Les petits objets comme surface de la dent unique peuvent être numérisés avec une précision encore plus élevée, qui permet au système d&#39;évaluer érosive et dent abrasif perte de surface. Le scanner de référence peut être utilisée pour mesurer les différences pour un grand nombre de domaines de recherche dentaires. Les différents niveaux de grossissement combinés avec une précision locale et générale de haut peuvent être utilisés pour évaluer les changements de dents ou des restaurations simples jusqu&#39;à des changements de arcade complète.

Figure 16A montre l&#39;analyse d&#39;un échantillon de gypse. Pour vérifier les paramètres de balayage optimal, contrôler la répétabilité de la matière. Une partie de l&#39;échantillon est analysé avec différents réglages de contraste et de luminosité et la répétabilité est vérifié après chaque balayage de trouver les réglages optimaux de balayage. Les surfaces métalliques montrent les meilleurs résultats avec un faible contraste 1.0 et matériaux de gypse ou de résine avec un co...

Watch this Scientific Journal Video about Accuracy in Dental Medicine, A New Way to Measure Trueness and Precision at JoVE.com

Accuracy in Dental Medicine, A New Way to Measure Trueness and Precision

La précision est une demande importante dans la médecine dentaire. Pour vérifier l&#39;exactitude, scanners de référence sont nécessaires. Cet article présente un nouveau scanner de référence avec une méthode de balayage ajusté à acquérir un large éventail de morphologies dentaires de haute justesse et précision.

Précision en médecine dentaire, une nouvelle façon de mesurer la justesse et fidélité

Reference scanners are used in dental medicine to verify a lot of procedures. The main interest is to verify impression methods as they serve as a base ...

accuracy-dental-medicine-new-way-to-measure-trueness

Research

JoVE Journal

Medicine

13.3K vues.  University of Zürich.  La précision est une demande importante dans la médecine dentaire. Pour vérifier l'exactitude, scanners de référence sont nécessaires. Cet article présente un nouveau scanner de référence avec une méthode de balayage ajusté à acquérir un large éventail de morphologies dentaires de haute justesse et précision. 

Vidéo: Accuracy in Dental Medicine, A New Way to Measure Trueness and Precision

Technique to Collect Fungiform (Taste) Papillae from Human Tongue

The sense of taste is critical for human life. It informs the body about the quality of food that will be potentially ingested and stimulates metabolic processes that prepare the alimentary canal for digestion. Steady progress is being made towards understanding the early biochemical and molecular events underlying taste transduction (for a review, Breslin and Spector, 20081). However, progress to date has largely resulted from animal models. Yet, since marked differences in receptor specificity and receptor density vary among species, human taste transduction will only be understood by using human taste tissue. Here we describe a biopsy technique to collect human fungiform papillae, visible as rounded pink anterior structures, about 0.5 mm in diameter that contain taste buds. These biopsied papillae are used for several purposes including the isolation of viable taste bud cells, in situ hybridization, immunohistochemistry and, through techniques of molecular biology, the identification of taste-specific novel proteins.

Technique to Collect Fungiform Taste Papillae from Human Tongue

Knowledge of molecular mechanisms underlying gustatory transduction has recently enjoyed significant advances, largely due to using animal models. However, the wide diversity in taste sensitivity and specificity among mammals warrants studies in human tissue. We describe a biopsy technique to collect living taste cells from the papillae on human tongue.

Technique pour collecter des fongiformes (goût) Papilles de la langue de l&#39;homme

The sense of taste is critical for human life. It informs the body about the quality of food that will be potentially ingested and stimulates metabolic ...

Recherche

Médecine

Construction of Vapor Chambers Used to Expose Mice to Alcohol During the Equivalent of all Three Trimesters of Human Development

Exposure to alcohol during development can result in a constellation of morphological and behavioral abnormalities that are collectively known as Fetal Alcohol Spectrum Disorders (FASDs). At the most severe end of the spectrum is Fetal Alcohol Syndrome (FAS), characterized by growth retardation, craniofacial dysmorphology, and neurobehavioral deficits. Studies with animal models, including rodents, have elucidated many molecular and cellular mechanisms involved in the pathophysiology of FASDs. Ethanol administration to pregnant rodents has been used to model human exposure during the first and second trimesters of pregnancy. Third trimester ethanol consumption in humans has been modeled using neonatal rodents. However, few rodent studies have characterized the effect of ethanol exposure during the equivalent to all three trimesters of human pregnancy, a pattern of exposure that is common in pregnant women. Here, we show how to build vapor chambers from readily obtainable materials that can each accommodate up to six standard mouse cages. We describe a vapor chamber paradigm that can be used to model exposure to ethanol, with minimal handling, during all three trimesters. Our studies demonstrate that pregnant dams developed significant metabolic tolerance to ethanol. However, neonatal mice did not develop metabolic tolerance and the number of fetuses, fetus weight, placenta weight, number of pups/litter, number of dead pups/litter, and pup weight were not significantly affected by ethanol exposure. An important advantage of this paradigm is its applicability to studies with genetically-modified mice. Additionally, this paradigm minimizes handling of animals, a major confound in fetal alcohol research.

We demonstrate the construction of alcohol vapor chambers using readily available materials that simultaneously house 6 mouse cages. We further describe their use in a mouse model of fetal alcohol exposure equivalent to all 3&#160;trimesters of human pregnancy. This paradigm exposes animals during gestation and postnatal days 1-12.

Construction des chambres de vapeur utilisée pour exposer des souris à l&#39;alcool pendant l&#39;équivalent de toutes trois trimestres de développement humain

Exposure to alcohol during development can result in a constellation of morphological and behavioral abnormalities that are collectively known as Fetal ...

Intrauterine Telemetry to Measure Mouse Contractile Pressure In Vivo

A complex integration of molecular and electrical signals is needed to transform a quiescent uterus into a contractile organ at the end of pregnancy. Despite the discovery of key regulators of uterine contractility, this process is still not fully understood. Transgenic mice provide an ideal model in which to study parturition. Previously, the only method to study uterine contractility in the mouse was ex vivo isometric tension recordings, which are suboptimal for several reasons. The uterus must be removed from its physiological environment, a limited time course of investigation is possible, and the mice must be sacrificed. The recent development of radiometric telemetry has allowed for longitudinal, real-time measurements of in vivo intrauterine pressure in mice. Here, the implantation of an intrauterine telemeter to measure pressure changes in the mouse uterus from mid-pregnancy until delivery is described. By comparing differences in pressures between wild type and transgenic mice, the physiological impact of a gene of interest can be elucidated. This technique should expedite the development of therapeutics used to treat myometrial disorders during pregnancy, including preterm labor.

Intrauterine Telemetry to Measure Mouse Contractile Pressure In Vivo

This manuscript provides a protocol for implanting telemeters in the mouse for the purpose of measuring intrauterine pressures during pregnancy.

Télémétrie intra-utérine mesurer souris contractile pression<em&gt; In Vivo</em

A complex integration of molecular and electrical signals is needed to transform a quiescent uterus into a contractile organ at the end of pregnancy. ...

Studying Pancreatic Cancer Stem Cell Characteristics for Developing New Treatment Strategies

Pancreatic ductal adenocarcinoma (PDAC) contains a subset of exclusively tumorigenic cancer stem cells (CSCs) which have been shown to drive tumor initiation, metastasis and resistance to radio- and chemotherapy. Here we describe a specific methodology for culturing primary human pancreatic CSCs as tumor spheres in anchorage-independent conditions. Cells are grown in serum-free, non-adherent conditions in order to enrich for CSCs while their more differentiated progenies do not survive and proliferate during the initial phase following seeding of single cells. This assay can be used to estimate the percentage of CSCs present in a population of tumor cells. Both size (which can range from 35 to 250 micrometers) and number of tumor spheres formed represents CSC activity harbored in either bulk populations of cultured cancer cells or freshly harvested and digested tumors 1,2. Using this assay, we recently found that metformin selectively ablates pancreatic CSCs; a finding that was subsequently further corroborated by demonstrating diminished expression of pluripotency-associated genes/surface markers and reduced in vivo tumorigenicity of metformin-treated cells. As the final step for preclinical development we treated mice bearing established tumors with metformin and found significantly prolonged survival. Clinical studies testing the use of metformin in patients with PDAC are currently underway (e.g., NCT01210911, NCT01167738, and NCT01488552). Mechanistically, we found that metformin induces a fatal energy crisis in CSCs by enhancing reactive oxygen species (ROS) production and reducing mitochondrial transmembrane potential. In contrast, non-CSCs were not eliminated by metformin treatment, but rather underwent reversible cell cycle arrest. Therefore, our study serves as a successful example for the potential of in vitro sphere formation as a screening tool to identify compounds that potentially target CSCs, but this technique will require further in vitro and in vivo validation to eliminate false discoveries.

Pancreatic cancer stem cells (CSCs) can be expanded in vitro using the anchorage-independent sphere culture technique, which represents a powerful tool to study CSC biology and can serve as the first step to develop novel CSC-targeting therapies. Here the methodology for expanding, analyzing and targeting of pancreatic CSCs is provided.

Etudier le cancer du pancréas Stem Cell Caractéristiques pour le développement de nouvelles stratégies de traitement

Pancreatic ductal adenocarcinoma (PDAC) contains a subset of exclusively tumorigenic cancer stem cells (CSCs) which have been shown to drive tumor ...

Boldness, Aggression, and Shoaling Assays for Zebrafish Behavioral Syndromes

A behavioral syndrome exists when specific behaviors interact under different contexts. Zebrafish have been test subjects in recent studies and it is important to standardize protocols to ensure proper analyses and interpretations. In our previous studies, we have measured boldness by monitoring a series of behaviors (time near surface, latency in transitions, number of transitions, and darts) in a 1.5 L trapezoidal tank. Likewise, we quantified aggression by observing bites, lateral displays, darts, and time near an inclined mirror in a rectangular 19 L tank. By dividing a 76 L tank into thirds, we also examined shoaling preferences. The shoaling assay is a highly customizable assay and can be tailored for specific hypotheses. However, protocols for this assay also must be standardized, yet flexible enough for customization. In previous studies, end chambers were either empty, contained 5 or 10 zebrafish, or 5 pearl danios (D. albolineatus). In the following manuscript, we present a detailed protocol and representative data that accompany successful applications of the protocol, which will allow for replication of behavioral syndrome experiments.

This manuscript describes the setup, implementation, and analysis of boldness, aggression, and shoaling in zebrafish and testing for the presence of a behavioral syndrome. A standardized approach for behavioral quantification will allow for easier comparison across studies. Modifications to this protocol are possible as each assay can be run individually.

Audace, Agression, et Essais de shoaling pour Zebrafish Behavioral Syndromes

A behavioral syndrome exists when specific behaviors interact under different contexts. Zebrafish have been test subjects in recent studies and it is ...

Open Tracheostomy Gastric Acid Aspiration Murine Model of Acute Lung Injury Results in Maximal Acute Nonlethal Lung Injury

Acid pneumonitis is a major cause of sterile acute lung injury (ALI) in humans. Acid pneumonitis spans the clinical spectrum from asymptomatic to acute respiratory distress syndrome (ARDS), characterized by neutrophilic alveolitis, and injury to both alveolar epithelium and vascular endothelium. Clinically, ARDS is defined by acute onset of hypoxemia, bilateral patchy pulmonary infiltrates and non-cardiogenic pulmonary edema. Human studies have provided us with valuable information about the physiological and inflammatory changes in the lung caused by ARDS, which has led to various hypotheses about the underling mechanisms. Unfortunately, difficulties determining the etiology of ARDS, as well as a wide range of pathophysiology have resulted in a lack of critical information that could be useful in developing therapeutic strategies.
Translational animal models are valuable when their pathogenesis and pathophysiology accurately reproduce a concept proven in both in vitro and clinical settings. Although large animal models (e.g., sheep) share characteristics of the anatomy of human trachea-bronchial tree, murine models provide a host of other advantages including: low cost; short reproductive cycle lending itself to greater data acquisition; a well understood immunologic system; and a well characterized genome leading to the availability of a variety of gene deletion and transgenic strains. A robust model of low pH induced ARDS requires a murine ALI that targets mainly the alveolar epithelium, secondarily the vascular endothelium, as well as the small airways leading to the alveoli. Furthermore, a reproducible injury with wide differences between different injurious and non-injurious insults is important.
The murine gastric acid aspiration model presented here using hydrochloric acid employs an open tracheostomy and recreates a pathogenic scenario that reproduces the low pH pneumonitis injury in humans. Additionally, this model can be used to examine interaction of a low pH insult with other pulmonary injurious entities (e.g., food particles, pathogenic bacteria).

This protocol induces acute lung injury in a mouse that has close fidelity to the pathogenesis of acid pneumonitis observed in humans. We generate a maximal acute nonlethal low pH lung injury and account for differences in rodent-human anatomic respiratory structure using an open tracheostomy coupled with circumferential pressure release.

Ouvrir trachéotomie acide gastrique Aspiration modèle murin de lésion pulmonaire aiguë Résultats dans Maximal aiguë non létaux Lung Injury

Acid pneumonitis is a major cause of sterile acute lung injury (ALI) in humans. Acid pneumonitis spans the clinical spectrum from asymptomatic to acute ...

Conducting Maximal and Submaximal Endurance Exercise Testing to Measure Physiological and Biological Responses to Acute Exercise in Humans

Regular physical activity has a positive effect on human health, but the mechanisms controlling these effects remain unclear. The physiologic and biologic responses to acute exercise are predominantly influenced by the duration and intensity of the exercise regimen. As exercise is increasingly thought of as a therapeutic treatment and/or diagnostic tool, it is important that standardizable methodologies be utilized to understand the variability and to increase the reproducibility of exercise outputs and measurements of responses to such regimens. To that end, we describe two different cycling exercise regimens that yield different physiologic outputs. In a maximal exercise test, exercise intensity is continually increased with a greater workload resulting in an increasing cardiopulmonary and metabolic response (heart rate, stroke volume, ventilation, oxygen consumption and carbon dioxide production). In contrast, during endurance exercise tests, the demand is increased from that at rest, but is raised to a fixed submaximal exercise intensity resulting in a cardiopulmonary and metabolic response that typically plateaus. Along with the protocols, we provide suggestions on measuring physiologic outputs that include, but are not limited to, heart rate, slow and forced vital capacity, gas exchange metrics, and blood pressure to enable the comparison of exercise outputs between studies. Biospecimens can then be sampled to assess cellular, protein, and/or gene expression responses. Overall, this approach can be easily adapted into both short- and long-term effects of two distinct exercise regimens.

To assess the influence of exercise intensity on physiologic and biologic responses, two different exercise testing protocols were utilized. Methods outlining exercise testing on a cycle ergometer as an incremental maximal oxygen consumption test and endurance, steady state submaximal endurance test are described.

Réalisation d’un exercice d’Endurance maximale et sous-maximale tests pour mesurer les réponses physiologiques et biologiques à un exercice aigu chez l’homme

Regular physical activity has a positive effect on human health, but the mechanisms controlling these effects remain unclear. The physiologic and biologic ...

Precision Implementation of Minimal Erythema Dose (MED) Testing to Assess Individual Variation in Human Inflammatory Response

Minimal erythema dose (MED) testing is frequently used in clinical settings for determining the smallest amount of ultraviolet (UV) irradiation necessary to produce erythema (inflammatory reddening) on the surface of the skin. In this context, the MED is regarded as a key factor in determining starting doses for UV phototherapy for common skin conditions such as psoriasis and eczema. In research settings, MED testing also has potential to be a powerful tool for assessing within- and between-persons variation in inflammatory responses. However, MED testing has not been widely adopted for use in research settings, likely owing to a lack of published guidelines, which is a barrier to obtaining reproducible results from this assay. Also, protocols and equipment for establishing MED vary widely, making it difficult to compare results across laboratories. Here, we describe a precise and reproducible method to induce and measure superficial erythema using newly designed protocols and methods that can easily be adapted to other equipment and laboratory environments. The method described here includes detail on procedures that will allow extrapolation of a standardized dosage schedule to other equipment so that this protocol can be adapted to any UV radiation source.

Precision Implementation of Minimal Erythema Dose MED Testing to Assess Individual Variation in Human Inflammatory Response

Minimal erythema dose (MED) testing is used to establish dosage schedules for ultraviolet radiation phototherapy. It can assess individual variation in inflammatory response but lacks methodology for achieving reproducible results. Here, we present a precision implementation of MED and demonstrate its ability to capture individual variation in inflammatory response.

Mise en œuvre avec précision des tests de dose minimale d'érythème (MED) pour évaluer la variation individuelle de la réponse inflammatoire humaine

Minimal erythema dose (MED) testing is frequently used in clinical settings for determining the smallest amount of ultraviolet (UV) irradiation necessary ...

Murine Precision-Cut Liver Slices as an Ex Vivo Model of Liver Biology

Understanding the mechanisms of liver injury, hepatic fibrosis, and cirrhosis that underlie chronic liver diseases (i.e., viral hepatitis, non-alcoholic fatty liver disease, metabolic liver disease, and liver cancer) requires experimental manipulation of animal models and in vitro cell cultures. Both techniques have limitations, such as the requirement of large numbers of animals for in vivo manipulation. However, in vitro cell cultures do not reproduce the structure and function of the multicellular hepatic environment. The use of precision-cut liver slices is a technique in which uniform slices of viable mouse liver are maintained in laboratory tissue culture for experimental manipulation. This technique occupies an experimental niche that exists between animal studies and in vitro cell culture methods. The presented protocol describes a straightforward and reliable method to isolate and culture precision-cut liver slices from mice. As an application of this technique, ex vivo liver slices are treated with bile acids to simulate cholestatic liver injury and ultimately assess the mechanisms of hepatic fibrogenesis.

This protocol provides a simple and reliable method for the production of viable precision-cut liver slices from mice. The ex vivo tissue samples can be maintained under laboratory tissue culture conditions for multiple days, providing a flexible model to examine liver pathobiology.

Murine Precision-Cut Liver Slices comme un modèle Ex Vivo de biologie du foie

Understanding the mechanisms of liver injury, hepatic fibrosis, and cirrhosis that underlie chronic liver diseases (i.e., viral hepatitis, non-alcoholic ...

Intraoperative Assessment of Resection Margins in Oral Cavity Cancer: This is the Way

The goal of head and neck oncological surgery is complete tumor resection with adequate resection margins while preserving acceptable function and appearance. For oral cavity squamous cell carcinoma (OCSCC), different studies showed that only 15%-26% of all resections are adequate. A major reason for the low number of adequate resections is the lack of information during surgery; the margin status is only available after the final histopathologic assessment, days after surgery.
The surgeons and pathologists at the Erasmus MC University Medical Center in Rotterdam started the implementation of specimen-driven intraoperative assessment of resection margins (IOARM) in 2013, which became the standard of care in 2015. This method enables the surgeon to turn an inadequate resection into an adequate resection by performing an additional resection during the initial surgery. Intraoperative assessment is supported by a relocation method procedure that allows accurate identification of inadequate margins (found on the specimen) in the wound bed.
The implementation of this protocol resulted in an improvement of adequate resections from 15%-40%. However, the specimen-driven IOARM is not widely adopted because grossing fresh tissue is counter-intuitive for pathologists. The fear exists that grossing fresh tissue will deteriorate the anatomical orientation, shape, and size of the specimen and therefore will affect the final histopathologic assessment. These possible negative effects are countered by the described protocol. Here, the protocol for specimen-driven IOARM is presented in detail, as performed at the institute.

The goal of this protocol is to provide a clear overview of specimen-driven intraoperative assessment of resection margins. It is encouraged to implement this protocol to improve patient care at other institutes.